Integrated Structural Modeling of Full-Length LRH-1 Reveals Inter-domain Interactions Contribute to Receptor Structure and Function

  • Structure. 2020 Jul 7;28(7):830-846.e9. doi: 10.1016/j.str.2020.04.020.
Corey D Seacrist  1 ,  Georg Kuenze  2 ,  Reece M Hoffmann  3 ,  Brandon E Moeller  3 ,  John E Burke  3 ,  Jens Meiler  4 ,  Raymond D Blind  5
Affiliations
  • 1. Department of Pharmacology, Vanderbilt University, Nashville, TN, USA; Center for Structural Biology, Vanderbilt University, Nashville, TN, USA.
  • 2. Department of Chemistry, Vanderbilt University, Nashville, TN, USA; Center for Structural Biology, Vanderbilt University, Nashville, TN, USA.
  • 3. Department of Biochemistry and Microbiology, University of Victoria, Victoria, BC, Canada.
  • 4. Department of Pharmacology, Vanderbilt University, Nashville, TN, USA; Department of Chemistry, Vanderbilt University, Nashville, TN, USA; Institute for Drug Discovery, Leipzig University, Leipzig, Germany; Center for Structural Biology, Vanderbilt University, Nashville, TN, USA. Electronic address: [email protected].
  • 5. Department of Pharmacology, Vanderbilt University, Nashville, TN, USA; Department of Medicine, Division of Diabetes Endocrinology and Metabolism, Vanderbilt University Medical Center, Nashville, TN, USA; Department of Biochemistry, Vanderbilt University, Nashville, TN, USA; Center for Structural Biology, Vanderbilt University, Nashville, TN, USA. Electronic address: [email protected].
PMID: 32433991 DOI: 10.1016/j.str.2020.04.020
Abstract

Liver receptor homolog-1 (LRH-1; NR5A2) is a nuclear receptor that regulates a diverse array of biological processes. In contrast to dimeric nuclear receptors, LRH-1 is an obligate monomer and contains a subtype-specific helix at the C terminus of the DNA-binding domain (DBD), termed FTZ-F1. Although detailed structural information is available for individual domains of LRH-1, it is unknown how these domains exist in the intact nuclear receptor. Here, we developed an integrated structural model of human full-length LRH-1 using a combination of HDX-MS, XL-MS, Rosetta computational docking, and SAXS. The model predicts the DBD FTZ-F1 helix directly interacts with ligand binding domain helix 2. We confirmed several Other predicted inter-domain interactions via structural and functional analyses. Comparison between the LRH-1/Dax-1 co-crystal structure and the integrated model predicted and confirmed Dax-1 co-repressor to modulate LRH-1 inter-domain dynamics. Together, these data support individual LRH-1 domains interacting to influence receptor structure and function.

Keywords
BS3; Dax1 Dax-1 Nr0b1; benzophenone artificial amino acid; disulfide crosslink mass spectrometry; hydrogen-deuterium exchange mass spectrometry; integrated structural modeling; nuclear lipids; nuclear phospholipid signaling; nuclear receptor lipidomics; small-angle X-ray scattering; β-catenin CTNNB1.